共刺激分子CD80和CD86在蠕虫保护性免疫及其Th2免疫应答中作用机制的研究

目的:探讨共刺激分子CD80和CD86在蠕虫保护性免疫及其Th2免疫应答中的作用机制。方法:采用巴西日圆线虫第3期幼虫感染BALB/c小鼠,在感染当天、感染后第3、7d分别腹腔注射大鼠抗CD80和/或抗CD86单克隆抗体,以阻断协同刺激信号。在感染当天,感染后第7、14d,计数小鼠外周血液的嗜酸性粒细胞,于感染后第14d计数小肠绒毛嗜酸性粒细胞;采用WatanabeN法测定IgE。结果:联合应用抗CD80和抗CD86两种单克隆抗体可导致成虫产卵量明显增加,成虫数量显著增多及排虫时间延迟。同时两种单克隆抗体也完全抑制了外周血液及小肠组织的嗜酸性粒细胞的增多并部分抑制了IgE水平的升高。而单独应用抗CD80或抗CD86McAb对保护性免疫及Th2型免疫应答反应均无明显影响。结论:在蠕虫保护性免疫和Th2型免疫应答反应中,T细胞的活化需要CD80和CD86两种共刺激分子的参与;而CD80或CD86单一刺激分子即可提供足够的共刺激信号。研究提示,人为调控CD80和CD86共刺激分子可能有助于对蠕虫病及Th2型免疫应答所介导的其它疾病的治疗。     

Phenotypes and spontaneous cell cytotoxicity of mononuclear cells from patients with seronegative spondyloarthropathies: Ankylosing spondylitis,psoriatic arthropathy and pauciarticular juvenile chronic arthritis — Analysis of mononuclear cells from peripheral blood,synovial fluid and synovial membranes

Summary T-cell subpopulations and natural killer (NK) cells from peripheral blood, synovial fluid and synovial membranes from patients with seronegative spondyloarthropathies were investigated. Thirty-four patients with ankylosing spondylitis, sixteen patients with psoriatic arthropathy and six patients with pauciarticular juvenile chronic arthritis were studied. All the patient groups had normal proportions of T4⁺ and T8⁺ cells as well as normal T4/T8 ratios in peripheral blood. In the synovial fluids the T4/T8 ratios were reduced in ankylosing spondylitis and psoriatic arthropathy (p<0.05). Although both the T4 and T8 subpopulations were reduced, the T4/T8 ratios in the synovial membranes of patients with these two disorders tended to be within the normal range of that of peripheral blood. Increased numbers of T-cells in the synovial fluid from patients with ankylosing spondylitis expressed class II MHC antigens. The natural killer cell activity was normal in peripheral blood and synovial fluids of patients with ankylosing spondylitis and psoriatic arthropathy while it tended to be reduced, although not significantly, in pauciarticular juvenile chronic arthritis. Synovial membranes were almost devoid of NK cell activity. The number of Leu 7⁺ cells were reduced in synovial fluid of patients with psoriatic arthropathy (p<0.04), but not as significantly as in the two other patient groups.     

CD26及半乳糖凝集素3免疫组织化学染色在甲状腺癌诊断中的应用

目的探讨CD26／DPPⅣ、半乳糖凝集素3免疫组织化学染色及其联合检测在甲状腺癌诊断中的应用价值。方法采用免疫组织化学EnVision二步法检测了114例良恶性甲状腺肿瘤组织中CD26／DPPⅣ及半乳糖凝集素3的表达。结果CD26／DPPⅣ及半乳糖凝集素3在正常甲状腺组织中无表达，在甲状腺腺瘤及滤泡癌中少有表达，在大多数甲状腺乳头状癌中呈不同程度的阳性表达。相对于甲状腺腺瘤而言，CD26／DPPⅣ诊断乳头状癌的敏感性、特异性、诊断准确率、阳性预测值、阴性预测值及kappa值分别为86．8％、97．2％、90．4％、98．3％．79．5％及0．80，半乳糖凝集素3分别为97．1％、91．7％、95．2％、95．7％、94．3％及0．89。结论CD26／DPPⅣ及半乳糖凝集素3均是甲状腺乳头状癌较为可靠的标志物，可以辅助常规的病理检查进行乳头状癌与腺瘤的鉴别诊断，它们在甲状腺滤泡癌诊断中的应用价值尚有待于进一步的研究。     

高迁移率族蛋白B1对大鼠脾脏树突状细胞表面共刺激分子表达的影响

目的观察高迁移率族蛋白B1(HMGB1)对树突状细胞(dendritic cells,DC)表面共刺激分子表达的影响,并对其机制进行初步探讨。方法分离正常Wistar大鼠脾脏DC后置于96孔培养板(1×10~5/孔),采用HMGB1刺激,观察HMGB1刺激与DC表面共刺激分子CD80、CD86和主要组织相容性复合物(MHC)Ⅱ表达的时间-效应关系及剂量-效应关系。结果HMGB1刺激后,DC表面共刺激分子CD80、CD86和MHCⅡ表达分别于24～72 h明显上调(P＜0．05,0．01),其中以作用48 h后DC表面共刺激分子表达上调尤为显著(P＜0．01)；0．1μg/ml、1μg/ml、10μg/ml的HMGB1刺激均可诱导DC表面共刺激分子CD80、CD86和MHCⅡ表达增强(P＜0．05,0．01),其中HMGB1的浓度在1μg/ml时,大鼠DC表面共刺激分子CD80、CD86和MHCⅡ的表达增强最明显(P＜0．01)。结论HMGB1能诱导DC表面共刺激分子表达增强,HMGB1可能是诱导DC成熟的免疫刺激信号。     

激素敏感型肾病综合征小儿的HLA

为了解中国汉族儿童激素敏感型肾病综合征与ＨＬＡ的关系，研究了３７名该病患儿的ＨＬＡ－Ａ，Ｂ，ＤＲ，ＤＱ抗原频率，ＨＬＡ－Ａ，Ｂ抗原与本病无关联，ＨＬＡ－ＤＲ７抗原频率（３７．８４％）较对照组（１１．２３％）有显著性增高（Ｐｃ＝１．７×１０－３），ＨＬＡ一ＤＱ抗原较对照组低（３２．４３％与６１．４０％，Ｐｃ＝７．７×１０－３），频复发或频反复病人与ＨＬＡ－ＤＲ９抗原相关（Ｐｃ＝２．９×ｌ０－２）。支持该病有免疫遗传基础的假说     

肺腺癌VEGF-C、VEGF-D的表达与MVD、MLVD及淋巴转移的关系

目的探讨肺腺癌中VEGF-C、VEGF-D与微淋巴管密度MLVD（VEGFR-3）、微血管密度MVD （CD34）及淋巴结转移之间的关系。方法免疫组化检测48例肺腺癌组织中VEGF-C、VEGF-D、MLVD、MVD蛋白的表达。结果VEGF-C、VEGF-D蛋白阳性率分别为70．8％（34／48例）、58．3％（28／48例）,肿瘤周边部位显著高于肿瘤中心部位,具有统计学意义,其表达与肿瘤分化程度无关,与肿瘤的TNM分期有关,Ⅲ～Ⅳ期显著高于Ⅰ～Ⅱ期。在VEGF-C蛋白阳性组,MVD高于阴性组（P=0．016）,MLVD显著高于阴性组（P=0．006）,淋巴结转移（P=0．042）增多;而VEGF-D蛋白阳性组与阴性组相比MVD无显著差异（P=0．943）, MLVD高于阴性组（P〈0．01）,淋巴结转移（P=0．012）增加。结论VEGF-C的表达与肺腺癌血管生成及淋巴管生成和淋巴结转移关系密切,而VEGF-D的表达只与淋巴管生成和淋巴结转移关系密切,与血管生成无关。     

Flow Cytometric Analysis of Lymphocyte Subsets of Mice Maintained on an Ethanol-Containing Liquid Diet

Alcoholic patients often have impaired immune function, yet little is known about the precise mechanism(s) of this impairment. We have previously shown that ethanol consumption by mice alters copolymer-specific humoral and cellular immune responses. In this study, we asked whether alcohol consumption by mice would phenotypically alter lymphocyte populations. Female C57BL/6 mice were fed a nutritionally complete liquid diet containing 35% ethanol-derived calories for up to 8 days. As controls, mice either were fed a liquid control diet that isocalorically substitutes sucrose for ethanol or remained on a standard solid diet and water ad libitum. Although mice fed ethanol-containing liquid or pair-fed control liquid diets have decreased numbers of spleen cells compared with solid diet controls, only the ethanol-containing diet allowed normally nonresponder C57BL/6 spleen cells to make antibody responses to the poly(Glu⁵⁰Tyr⁵⁰) synthetic copolymer antigen. Flow cytometric analysis of splenic lymphocyte populations of mice on the ethanol-containing diet shows an increase in the relative proportion of T-lymphocytes as compared with mice on either solid or liquid control diets. No such change is seen for either B-cell or natural killer cell populations in these same mice. Both liquid control and liquid ethanol diets caused a slight decrease in the CD4:CD8 ratios of splenic T-lymphocytes. We see the relative percentage of T-cells bearing the αβ-cell receptor (TcR) increases in the spleens of liquid ethanol diet mice; a smaller increase TcRαβ usage is seen in the spleens of liquid control mice, compared with solid diet mice. Flow cytometric analysis shows that little, if any, difference exists in TcRγδ expression between the liquid ethanol and either the liquid control or solid diet groups. Preliminary analysis of TcRαβ subsets suggest that ethanol increases the percentage of T-cells expressing Vβ5 and Vβ8, and decreases the percentage of Vβ11 expressing cells. These findings suggest that, in addition to modifying the immune response, ethanol alters the phenotypic expression of lymphocyte subsets.     

A novel nucleotide-containing antigen for human blood γδ T lymphocytes

The stimulation of human γδ T cells by mycobacteria occurs through recognition of four distinct nonpeptide phosphorylated antigens termed TUBag1–4. Among these latter, TUBag4 has already been biochemically characterized as a γ-X derivative of 5′-deoxythymidine triphosphate (Constant, P., Davodeau, F., Peyrat, M. A., Poquet, Y., Puzo, G., Bonneville, M. and Fournié, J.-J., Science 1994. 264: 267). However, despite chemical synthesis of weakly stimulatory nucleotide-containing analogs, these mycobacterial compounds remained the sole nucleotide-containing antigens actually isolated from natural sources. Here, we present the complete isolation of the TUBag3 antigen from Mycobacterium fortuitum and demonstrate that this nonpeptide molecule contains a 5′-UTP nucleotide moiety. On selected Vγ9/Vδ2 clones, T cell responses can be triggered with nanomolar concentrations of TUBag3. Like crude mycobacterial extracts, this purified nucleotide conjugate elicits a strong polyclonal response of γδ PBL from healthy donors. Furthermore, we present evidence that this compound is distinct from the recently synthesized γ-isopentenyl 5′-UTP, a nucleotide conjugate of isopentenyl pyrophosphate that was found to be stimulatory for human γδ T cells (Tanaka, Y., Morita, C. T., Tanaka, Y., Nieves, E., Brenner, M. B. and Bloom, B. R., Nature 1995. 375: 155). Since it appears that both mycobacterial nucleotide antigens are molecules structurally related to peculiar precursors of nucleic acid synthesis, we propose that TUBag-reactive T cells might be specifically devoted to surveillance of proliferating cells.     

Immunohistochemical expression of MAM-3 and MAM-6 antigens in salivary gland tumours

Summary MAM-3 and MAM-6 antigens of human milk fat globule membrane were detected immunohistochemically in 93 cases of salivary gland tumours as well as in normal glands. The antigens were visualized in 10% formalin-fixed paraffin sections. MAM-3 (MoAbs 115G3, 67D11) antigen was distributed in intercalated and striated duct cells of the normal salivary glands, and in luminal tumour cells and squamous metaplastic cells of pleomorphic adenomas. In pleomorphic adenomas the frequency of positive staining with MoAb 67D11 (54/67; 80.6%) was higher than that with MoAb 115G3 (36/67; 53.7%). MAM-6 (MoAbs 115D8, 115F5) antigen was expressed in luminal and lateral borders of serous acinar cells and ductal of the normal glands, and also in luminal borders of tubulo-ductal and glandular structures of salivary gland tumours. Ductal basal cells were characterized by existence of positive staining for MAM-6 antigen, in adenolymphomas MAM-6 antigen was restricted to the basal tumour cells. Some mucous cells of mucoepidermoid tumours were stained specifically with MoAb 115G3, and epidermoid cells of mucoepidermoid carcinomas manifested MAM-6 antigen staining. Immunohistochemical localization of MAM-6 antigen resembled that of epithelial membrane antigen (EMA) detected with MoAb.     

Anti-high endothelial venule monoclonal antibody HECA-452 recognizes plasmacytoid T cells and delineates an ”extranodular“ compartment in the reactive lymph node

So-called plasmacytoid T cells represent a subset of monocyte related cells, which share with endothelium the CD36⁺CD11b⁻ (OKM5⁺OKM1⁻) phenotype. The reactivity of plasmacytoid T cells with rat monoclonal antibody HECA-452, highly specific for high endothelial venules, was analyzed in reactive lymph nodes. In all cases, HECA-452 not only labelled the endothelium of high endothelial venules, but also strongly reacted with singular and clustered plasmacytoid T cells. The HECA-452 positivity for high endothelial venules and plasmacytoid T cells visualized a lymph node compartment extending from the subcapsular sinus to the corticomedullary junction. This compartment surrounded the composite nodule and was designated the ”extranodular“ compartment. The cooccurrence of plasmacytoid T cells and high endothelial venules in this extranodular compartment, together with their immunophenotypical similarities, may be indicative of functional co-operations.